Integrin alpha 5 Recombinant Rabbit Monoclonal Antibody [JJ08-94]
cat.: ET1701-58
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Monkey, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: JJ08-94
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 115 kDa
Isotype: IgG
Immunogen: Synthetic peptide within C-terminal human Integrin alpha 5.
Positive control: HeLa cell lysate, A549 cell lysate, U-937 cell lysate, NIH/3T3 cell lysate, COS-1 cell lysate, Mouse liver tissue lysate, Rat liver tissue lysate, NIH/3T3, human kidney tissue, human placenta tissue, human tonsil tissue, mouse endometrium tissue, Hela, Jurkat.
Subcellular location: Membrane, Cell junction, Cell surface.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:1,000-1:2,000
1:50-1:200
1:50-1:200
1:200-1:1,000
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: P08648 Human | P11688 Mouse
Unigene: 100796 Rat
Alternative names: CD49 antigen-like family member E CD49e Fibronectin receptor subunit alpha Fibronectin receptor, alpha subunit FNRA Integrin alpha 5 (fibronectin receptor alpha) Integrin alpha-5 Integrin alpha-5 light chain Integrin alpha-F Integrin, alpha 5 (fibronectin receptor, alpha polypeptide) ITA5_HUMAN Itga5 Very late activation protein 5, alpha subunit VLA-5 VLA5 VLA5A
Images
ET1701-58_1.jpg Fig1: Western blot analysis of Integrin alpha 5 on different lysates with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A549 cell lysate
Lane 3: U-937 cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: COS-1 cell lysate
Lane 6: Mouse liver tissue lysate
Lane 7: Rat liver tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 115 kDa
Observed band size: 140/19 kDa

Exposure time: 1 minute 2 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-58) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1701-58_2.jpg Fig2: Immunocytochemistry analysis of NIH/3T3 cells labeling Integrin alpha 5 with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) at 1/200 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1701-58_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-58) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1701-58_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-58) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1701-58_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-58) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1701-58_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse endometrium tissue with Rabbit anti-Integrin alpha 5 antibody (ET1701-58) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-58) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1701-58_7.jpg Fig7: ICC staining of Integrin alpha 5 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-58, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1701-58_8.jpg Fig8: Flow cytometric analysis of Integrin alpha 5 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1701-58, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.